Crataegus pentagyna willd. Fruits, leaves and roots: phytochemicals, antioxidant and antimicrobial potentials

Background The hawthorn has recently been used as a popular herbal medicine in food applications and phytotherapy, especially for the cardiovascular system. Methods In this study, phytochemicals were evaluated by LC-ESI-MS, GC-MS, and biological activity, including antioxidant (DPPH test) and antibacterial (broth dilution assay), in different extracts of Crataegus pentagyna fruit, leaf, and root. Results Globally, 49 phenolics were tentatively identified using HPLC-ESI-MS/MS in the hydro-methanolic extract of the fruit (major apigenin, caffeoylquinic acid derivative, and 4-O-(3′-O-glucopyranosyl)-caffeoyl quinic acid), 42 in the leaf (major salicylic acid, naringenin-6-C-glucoside, and naringin), and 33 in the root (major naringenin-7-O-neohesperidoside, isovitexin-2″-O-rhamnoside, and 4-O-(3′-O-glucopyranosyl)-caffeoyl quinic acid). The major group compounds analyzed by GC-MS in petroleum ether extracts were hydrocarbons (63.80%) and fatty acids and their derivatives (11.77%) in fruit, hydrocarbons (49.20%) and fatty acids and their derivatives (13.85%) in leaf, and hydrocarbons (53.96%) and terpenes (13.06%) in root. All samples exhibited promising phytochemical profile (total phenol, flavonoid, phenolic acid, and anthocyanin), antioxidant and antibacterial capacities, especially in hydro-methanolic extract of fruit (210.22 ± 0.44 mg GAE/g DE; 79.93 ± 0.54 mg QE/g DE; 194.64 ± 0.32 mg CAE/g DE; 85.37 ± 0.13 mg cyanidin 3-glucoside/100 g FW; DPPH: 15.43 ± 0.65 µg/mL; MIC: 0.15–0.62 µg/mL; and MBC: 0.62–1.25 mg/mL), followed by the leaf and root extracts, respectively. The PCA and heatmap analysis results distinguished metabolite profile differences for samples. Conclusion The results of the present work provide scientific support for C. pentagyna as antimicrobial agents and natural antioxidants in human health and food preservation. Supplementary Information The online version contains supplementary material available at 10.1186/s12906-024-04430-4.


Introduction
Oxidative stress caused by overproduction of free radicals and reactive oxygen species that results in the development of several diseases such as diabetes, alzheimer's disease, parkinson's disease and cardiovascular conditions such as atherosclerosis, stroke and high blood pressure [1,2].Also, infectious diseases such as measles, flu, HIV, COVID-19, strep throat and salmonella are disorders caused by organisms such as viruses, bacteria, fungi or parasites [3].Infectious diseases resistance to antibiotics are 3rd common cause of death worldwide after cardiovascular diseases [4].So, there is a continuous need for research and investment in the field of new drugs and food preservatives with lower toxicity and higher efficacy.Recently, pharmaceutical and food industries have utilized medicinal plants as sources of bioactive substances, particularly phenolic compounds with a wide range of pharmacological activities [5].Most research found that phenolic compounds specially flavonoids exert antibacterial activity via damaging microbial cell membranes and inhibiting microbial enzymes and gene expression [6] and antioxidant activity via decreasing enzymatic activity of oxidases [7].Hawthorn, the common name for more than a thousand species of plants in the genus Crataegus and family Rosaceae (subfamily Maloideae).The genus of Crataegus is primarily found in Asia, Europe, and North America and is recommended by the World Health Organization as a medicinal and food ingredient in several countries.This genus is native to northern temperate regions and consists of 15-to 18-foot-tall trees and deciduous shrubs [8].Flowers, fruits, leaves, stems, and roots of Crataegus species have been recommended in modern and traditional medicine as cardiotonic, antispasmodic, diuretic, anti-atherosclerotic, and hypotensive agents [9,10].In terms of biological activity, proanthocyanidins and flavonoid glycosides are the most important compounds in hawthorn.In the leaves, flowers, and fruits of hawthorn, known phenolic compounds, such as quercetin, isoquercetin, rutin, hyperoside, epicatechin, chlorogenic acid, and protocatechuic acid, may be excellent sources of antioxidants [9][10][11].Variations in genetics, maturity of plant organs, collection regions, processing methods, and preharvest and postharvest environmental conditions may influence the chemical compound content of plant organs [12,13].In Iran's flora, Crataegus pentagyna subsp.elburensis is the most common cultivar of hawthorn.This plant is primarily utilized as an antiarrhythmic and cardiovascular disease preventative.The presence of polyphenols, such as flavonoids, phenolic acids, and proanthocyanidins, in the species may be primarily responsible for these effects [14].Earlier phytochemical investigations of C. pentagyna from different origins have revealed the identification of different compounds such as gallic acid, caffeic acid, and chlorogenic acid in fruit, pulp and seed of Iranian species [15,16]; hyperoside, rutin, isoquercitrin, sexangularetin-3-Oglucoside, isoorientin, isoorientin-2-O-rhamnoside, isovitexin, orientin, orientin-2-O-rhamnoside, vitexin, and vitexin-2-O-rhamnoside in leaves of Austrian species [17]; coumaric acid, chlorogenic acid, caffeic acid, ferulic acid, quercetin 3-O-glucoside (isoquercetin), quercetin, quercetin 3-O-rutinoside (rutin), (-)-epicatechin, kaempferol 3-O-glucoside, hyperoside, apigenin, cyanidin 3-O-glucoside, luteolin and procyanidins B1 and B2 in fruits, flowers and leaves of Serbian species [18]; and a number of flavonoid aglycones, flavonoid O-and C-glycosides, organic and phenolic acids and proanthocyanidins in leaf, flower and fruit of Romanian species [19].Previous studies showed that these phytochemicals have been linked to the health-promoting effects of this species, including cardiovascular system influence, antioxidant, anti-cancer, antimicrobial, anti-inflammatory and antihypercholesterolemic activities [9,20].As an effective method with high sensitivity and resolution, liquid chromatography coupled with electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) is widely used for plant metabolomics analyses and species discrimination.To our knowledge, there are no reports comparing the chemical profile and biological activities of different morphological parts of C. pentagyna.We investigated the chemical composition, total phenol, total flavonoid, total phenolic acid, total anthocyanin, antioxidant, and antibacterial activities of C. pentagyna fruits, leaves, and roots collected in Golestan province of Iran.Using gas chromatography coupled with mass spectrometry (GC-MS) and LC-ESI-MS/MS techniques, the chemical profile of petroleum ether and hydro-methanolic extracts was determined.

Plant samples
The essential factors for growth of plant are water, sunlight, heat, and topographical conditions in the planting region.Research has displayed that the optimum climate conditions for C. pentagyna growing involve 300-400 mm of annual precipitation, 8-12.5 °C of annual mean temperature, 2300-3200 °C of annual cumulative temperature, 2300 h of annual sunshine, and soil pH from 6.0 to 7.0.The highest growth rate occurs in the orchard between the ages of one and five years.Identification of optimal harvesting season is necessary to ensure contents of desired bioactive compounds.Therefore, around 500 g of fruits, leaves, and roots without damage of Crataegus pentagyna Willd.from one plant population were collected from Farsian, Galikesh, Golestan province, Iran (37°16'01.6"N55°26'09.5"E) in September 2020 (early maturity stage) at the highest content of phenolic compounds [16,18].Samples was verified by Dr. Ali Satarian and the voucher number (803,892) was deposited in the Herbarium of Gonbad Kavous University, Gonbad, Iran.

Preparation of plant extracts
Fresh fruits, leaves, and roots were air-dried and ground separately using a mortar.One gram of defatted powder with petroleum ether (3 × 20 mL) was extracted thrice separately in the dark for two hours with water-methanol and water-ethanol 80% (3 × 20 mL).Under reduced pressure, the filtrate extracts were evaporated at 40 °C on a rotary evaporator (Heidolph, Laborota 4000, Schwabach, Germany) to dryness and then freeze-dried.The dried extracts were stored at − 20 °C in the dark until further examination.In addition, the yield of constituents and the weight of dried extracts were determined [18].

Estimation of phenolic profile Total phenolic content (TPC)
The TPC of the extracts was determined using a modified version of Singleton and Rossi's (1965) method [21], with gallic acid as the standard.To 500 µL of diluted samples, a mixture of 2.5 mL of Folin-Ciocalteu (0.2 N) reagent and 2 mL of Na 2 CO 3 (75 g/L) was added.The absorbance of the samples was measured at 765 nm following 30 min of incubation at 45 °C.The results were given in milligrams of gallic acid per gram of dry extract (mg GAE/g DE).

Total flavonoid content (TFC)
Using a modified aluminum chloride colorimetric method [22], the TFC content of the samples was determined.In total, 0.5 mL of samples was combined with 0.1 mL of sodium acetate (1 M) and 0.1 mL of AlCl 3 (10%) and incubated for 30 min at room temperature.At 415 nm, the absorbance of the reaction mixture was measured.The total flavonoid concentration was calculated as mg of quercetin equivalents (QE) per gram of dry extract.

Determination of total phenolic acid content (TPAC)
The total phenolic acid content of the extracts was determined by spectrophotometry at 474 nm [23].A total of 1 mL of each extract was combined with 10 g of sodium nitrite and 10 g of sodium molybdate diluted in 100 mL of water, 2 mL of HCl (0.5 M), 3 mL of water, and 2 mL of NaOH (8.5% w/v) in this procedure.The results were reported in terms of mg of caffeic acid (CAE) per g of dry extract (mg CAE/g DE).

Determination of total anthocyanin content
The total anthocyanin concentration was determined using a method previously described [24].At 530 nm, the absorbance of diluted extracts with 1% HCl in methanol (5:95, v/v) was measured.Using the following equation, the values were expressed as mg malvidin-3-glucoside equivalents per g of dry extract.
The results were expressed in milligrams of cyanidin-3-glucoside per 100 g of fresh weight (mg c3g/100 g FW).

HPLC-PAD analysis
An analytical technique A KNAUER liquid chromatograph system with a photodiode array detector (Smartline PDA 2600) and a quaternary pump (Smartline Pump 1000) has been developed.The dissolved extracts in methanol: water 7:3 (approximately 2 mg/mL) were subjected to a gradient method on a 150 mm length × 4.6 mm inner-diameter C18 amide column (Varian, Darmstadt, Germany) at a flow rate of 1.0 mL/min and injection volume of 20 µL (water as solvent A and methanol as solvent B, including 0.05% trifluoroacetic acid).The elution gradient was 0-10 min with 10% solvent B, 10-35 min with 10-100% B, 35-45 min with 100% B, 45-50 min with 100-10% B, and 50-55 min with 10% B. The system was managed using the software EZ Chrom Elite.For testing the system's dependability, 4-hydroxymethyl benzoate, uracil, benzophenone, and 4-hydroxy ethyl benzoate were injected into the HPLC [25,26].

LC-ESI-MS/MS analysis of phenolic compounds
Using a Waters Alliance 2695 HPLC system coupled to a micro mass quattro micro API mass spectrometer with an ion source, the flavonoids and other phenolics compositions of hydro-methanolic extracts from the fruit, leaf, and root of C. pentagyna were determined (ESI).The separation was carried out using a Supelco C18 (15 mm×2.1 mm×3 μm) column with a flow rate of 0.2 mL/min and an injection volume of 10 µL.Using a gradient method (acetonitrile + 0.1% formic acid as solvent A and water + 0.1% formic acid as solvent B), the samples were eluted.The elution gradient was 0-5 min with 10% solvent A, 5-10 min with 10-50% A, 10-16 min with 50% A, 16-20 min with 50-90% A, 20-24 min with 90% A, 24-26 min with 90-10% B, and 26-30 min with 10% A. The mass spectra were acquired using negative ionization and a range of 50 to 2000 m/z.The detection of peaks at 310 °C probe temperature and 3.5 kV probe voltage were monitored.The LC-MS data were analyzed using MZmine version 2.35 software [27].

GC-MS analysis of essential compounds
Experiments were performed using an Agilent 6890 A mass spectrometer and an Agilent 5973 gas chromatograph.A total of 1 mL of the diluted extracts was injected into the HP 5 MS column with a length of 30 m, an inner diameter of 0.25 mm, and a thickness of 0.25 μm.As a carrier gas, helium at a flow rate of 1.0 mL/min was utilized.The column temperature was set at 50 °C for 3 min before increasing to 280 °C at a rate of 4 °C/min.The temperature of the GC injector and MS transfer line were set to 250 and 230 °C, respectively.MS detection utilized an ionization energy of 70 eV and a mass range of 50-550 m/z.By comparing their mass fragmentation patterns and retention times with the Wiley 7.0 and NIST libraries, the compound profiles were identified in petroleum ether extracts [28,29].

Antioxidant activit DPPH radical-scavenging activity
Modifications were made to the previously reported method [30] to measure antioxidant activity.Briefly, 3 mL of diluted samples with concentrations of 10 to 200 µg/ mL were mixed with 1 mL of 100 M methanolic DPPH solution.The absorbance of the mixture was measured at room temperature at 517 nm.DPPH radical scavenging capacity was calculated using the following formula: Where, A 0 is the absorbance of the blank (without extracts) and A t is the absorbance of the extracts.
The antioxidant capacity of the samples was expressed as IC 50 .Using the regression equation, the IC 50 values were determined as the concentrations of extracts that inhibit 50% of DPPH radicals.In addition, BHT served as a positive control.

Estimation of antibacterial activity
In our laboratory, a Gram-negative (Escherichia coli ATCC 25,922) and a Gram-positive (Staphylococcus aureus ATCC 9144) bacteria were stored at 4 °C and used in this study.Iranian Research Organization for Science and Technology provided the examined bacteria.The microdilution broth technique [31] was used to evaluate the antimicrobial activity of C. pentagyna extracts.Twofold serial dilutions of extracts in Muller Hinton broth (MHB) were prepared in a microtiter ranging from 0.078 to 20 mg/mL, and one million colony-forming units per milliliter were added to each well.Gentamicin and MHB were used as positive and negative controls, respectively.After 24 h of incubation at 35 °C, MIC was defined as the lowest concentration of the extracts with no visible bacterial growth.Also, MBC was determined as the lowest concentration of the extracts that was resulted in bacterial death.

Statistics
Each experiment was performed three times under the same conditions.The results of each test and analysis were recorded as means and standard deviation.Oneway ANOVA test was used to compare variance analyses, and the SAS software (least significant difference (LSD) test) was utilized to compare means (p < 0.05).The chromatograms of fruits, leaves and roots of C. pentagyna were pre-processed using MZmine analysis software package.To perform further statistical analyses was performed through SIMCA software (version 14.1 Umetrics AB, Umea, Sweden), MZmine exported the peak list data for each sample, which included the average retention time (RT), m/z values and peak intensity (height).The exported data were subsequently mean-centered and Pareto-scaled prior to multivariate statistical analysis to enhance the contribution from medium-sized features without inflating the noise from quiet areas of the chromatogram.Principal Component Analysis (PCA) and related biplot and loading plot was used for multidimensional data to identify metabolite differences between samples.Furthermore, heatmap analysis was performed using import-exciting clusters to better show differences between metabolites.

Results and discussion
C. pentagyna extraction yield, quantitative evaluation of phenolic profile, and antioxidant power.
The chemical components of the fruit, leaf, and root of C. pentagyna were fractionated using ether petroleum, 80% aqueous methanol, and ethanol as extraction solvents.The yield of extracts varied between 2.4% and 9.6% for methanolic extracts, between 2.1% and 8.3% for ethanolic extracts, and between 0.9% and 3.6% for petroleum ether extracts.Due to the hydroxyl (-OH) and methoxy (-OCH 3 ) groups in their molecular structures [32], phenolic compounds possess the ability to scavenge free radicals.The phenolic content of hawthorn varies by cultivar, species, geographical location, harvest time, method of chemical determination of phytochemicals, and extraction preparation conditions [33].There are few studies on the phytochemical content of C. pentagyna compounds.In addition, there have been no reports of hawthorn root.In this study, the phytochemical content (phenol, flavonoid, phenolic acid, and anthocyanin) and antioxidant capacity (DPPH scavenging) of hydro-methanolic and hydro-ethanolic extracts of fruit, leaf, and root were evaluated and compared (Table 1).The results indicated that hydro-methanolic extracts contain more phytochemicals and have more antioxidant capacity than ethanol extracts.Fruit extract contained the highest concentrations of phenolic (210.22 ± 0.44 mg GAE/g DE), total flavonoid (79.93 ± 0.54 mg QE/g DE), total phenolic acid (194.64 ± 0.32 mg CAE/g DE), and total anthocyanin (85.37 ± 0.13 mg cyanidin 3-glucoside/100 g FW) among the tested extracts, followed by leaf and root extracts.The total content of phenol and flavonoid in methanolic extracts of fruit from various regions ranged from 69.12 to 186.72 mg GAE/g and 1.6 to 85.31 mg QE/g dry weight plant, respectively, as reported by other researchers [15,32,[34][35][36].In a different study, the TPC and TFC concentrations in methanolic leaf extracts were 206.94GAE/g and 57.08 mg (+)-catechin/g, respectively [37,38].There was a strong correlation between TPC and DPPH reduction.The phenolic and flavonoid content of fruit, leaf, and root extracts increases their antioxidant activity.The fruit extract with the highest phenolic and flavonoid content exhibited the highest DPPH radical scavenging capacity (IC 50 = 15.43 ± 0.65 g/mL), followed by leaf and root extracts (IC 50 = 34.67 ± 0.14 g/mL and 60.72 ± 0.32 g/ mL, respectively).Moreover, the fruit and leaf extracts exhibited potent activity comparable to the positive control butylate hydroxytoluene (BHT) (IC 50 = 49.02± 0.2 g/ mL), whereas the root extract was less active.Antioxidant activity of fruit and leaf extracts of C. pentaegyna from different regions have been already investigated.For the methanolic fraction of fruit, the IC 50 values for DPPH radical scavenging activity range from 17.48 to 341.29 g/ mL [32,34,36,39].Moreover, the DPPH inhibition of C. pentagyna leaf was quantified as 5708 g/mL in hydroacetonic extract and 2.34 M TE/g (micromoles of Trolox equivalents) in ethanolic extract, respectively [18,37,40].

Flavonoids
Flavonoids containing two benzene rings and one oxygenated ring were the most abundant phenols found in hawthorn in this study.According to Table 2

Identification of catechins, proanthocyanidins, and their derivatives
There are two major categories of procyanidins (A-type and B-type).(Epi)catechin units linked through C4 to C8 or C4 to C6 are called B-type procyanidins, while those with an additional bond (C2-O-C7) are called A-type.

Phenolic acids
Free and conjugated phenolic acids are classified into two groups: hydroxybenzoic acids and hydroxycinnamic acids.Increasing interest in the profile of phenolic acids is due to their possible health benefits and antioxidant activity.In mass spectrometry, phenolic acids and their glycoside derivatives are distinguished by the loss of an ion at m/z -162 Da (glucose or galactose), followed by the loss of ions at m/z -18 Da (hydroxyl), -15 Da (methyl), and − 44 Da (carbon dioxide).In this study, a total of 16 phenolic acids were identified.

Identification of hydroxybenzoic acids and their glycosidic derivatives
Compound 10 and 12 were identified as protocatechuic acid and salicylic acid.Also, compounds 13 and 14 can be tentatively assigned as hydroxybenzoic acid derivatives.Previous studies have identified compound 12 from fruits of C. monogyna; compound 10 from fruits of C. germanica and fruits, leaves and flowers of C. pentagyna [19,77]; 3-hydroxybenzoic acid and 4-hydroxybenzoic acid from fruits of C. germanica [45,56,68]; and hydroxybenzoic acid, hydroxybenzoic acid hexoside from fruits, leaves and flowers of C. pentagyna [19].
Comparison between phenolic compounds in different parts of C. pentagyna based on ion intensity.

Antimicrobial activities
Table 4 provides a summary of the antibacterial activities of petroleum ether and hydro-methanolic extracts of C. pentagyna fruit, leaf, and root against two pathogenic bacteria (Staphylococcus aureus and Escherichia coli).All examined extracts inhibited bacterial growth with MIC and MBC values ranging from 0.15 to 5.12 mg/ mL and 0.15 to 10.12 mg/mL, respectively.The activity of the petroleum ether extract of leaf was higher than that of the root and fruit extracts (MICs 1.25-5 mg/mL).The presence of terpenes and flavonoids explains the high antibacterial activity in petroleum ether and hydro-methanol extracts, respectively [83]. .few studies have been conducted on the crude extracts of C. pentagyna to date.Salmanian et al. [16] examined the antimicrobial activity of seed and pulp extracts against four clinical pathogens.These extracts inhibited bacterial strains, with MICs and MBCs ranging between 2.5 and 40 mg/mL and 5 and > 40 mg/mL, respectively.In the study of Safapour et al. [84], C. pentagyna fruit extract demonstrated potent antibacterial activity, especially against Gram-negative bacteria.In a separate study, fruit acetonic extract exhibited the highest antibacterial activity against Bacillus subtilis (MBC = 2.5 mg/mL) [85].These antibacterial effects may be attributable to the flavonoid content of   the extracts, consistent with our findings.Terpenoids are known for their antibacterial effect and aromatic qualities.Non-polar constituents are more soluble in nonpolar solvents and polar constituents are more soluble in polar solvents, so there was a different antibacterial effect between solvents.Polar and non-polar solvents have high capacity to dissolve active antimicrobial compounds than the medium polar solvent.Terpenoids are fat soluble, so these phytochemicals can be attracted to the petroleum ether solvent as non-polar solvent [86].

Principle component analysis (PCA) and heatmap analysis
The PCA model as the most common multivariate data analysis is an unsupervised method to reduce the dimensionality of huge multivariate data sets.As shown in Fig. 3 samples (CEF: fruit, CEL: leaf and CER: root) revealed a clear difference in the PCA model, indicating differences in metabolic fingerprints of samples.The resulting values of R2X(cum) and Q2(cum) of 0.96 and 0.98, respectively, indicating a good fitness, discrimination, and predictability of the PCA model.The loading plot (Fig. 4) and bipolot (Fig. 5) are useful for identifying the variable responsible for similarities and differences between samples.The assignment of these variables led to the identification of compounds that are responsible for separation in the score plot.derivative for fruir and root; and coumaroylquinic acid derivative, sinapic acid, coumaric acid, vitexin-O-rhamnoside derivative, naringenin-7-O-neohesperidoside, naringenin and procyanidin pentamer-hexoside for leaf and root.Also, caffeoylquinic acid derivative, coumaroylquinic acid derivative, protocatechuic acid, caffeic acid, hydroxybenzoic acid derivative, ferulic acid, orientin glycoside, vitexin-O-glucoside derivative, quercetin-O-glycoside derivative, orientin, 8-methoxykaempferol and kaempferol-3-O-rutinoside were found only in fruit; coumaroylquinic acid derivative, eriodictyol-7-glucuronide and procyanidin B3 7-glucoside in leaf; and caffeoylquinic acid derivative and procyanidin tetramerhexoside in root.The heatmap analysis was presented in Fig. S2 to better show differences between metabolites.
(Supplementary materials).The metabolites of Table 2 were displayed in the heat map as variables 1-62.The major metabolites based on the relative abundance in heat map included some phenolic compounds including variables 1,3,

Conclusions
In this study, the compound profiles in petroleum ether and hydro-methanolic extracts of C. pentagyna fruit, leaf, and root tentatively were identified and characterized.In addition, the chemical profile of hawthorn root was discovered for the first time.The fruit hydro-methanolic extract exhibited the highest levels of antioxidant and antibacterial activity, followed by the leaf and root extracts.Antibacterial and antioxidant activities of C. pentagyna extracts were attributed to the major phenolics and terpenes detected by HPLC-MS/MS and GC-MS.
Using LC-ESI-MS, it was possible to characterize 62 compounds including mainly flavone apigenin, phenolic acid salicylic acid and flavanone naringin in fruit, leaf and root, respectively.Also, bioactive compounds such as alkane nonacosane in fruit and leaf extracts and triterpene squalene in root extract were identified using GC-MS as major components.The results of the PCA and heatmap analysis distinguished metabolite profile differences for fruit, leaf and root samples into welldefined groups.To confirm their potential as phytotherapeutic agents, it is suggested that further research should be conducted, including the purification of the , the aerial parts and roots of C. pentagyna contain 63 phenolic compounds.In mass spectrometry, all O-glycosides, including glucose or galactose (162 Daltons), rhamnose (146 Daltons), pentose-xylose or arabinose (132 Daltons), and disaccharide structures-rutinose or neohesperidose, lost their sugar moiety (308 Daltons).Due to cross-ring cleavages of sugar residues, C-glycosides exhibited fragments at m/z [M-H-18] −− , [M-H-60] − , [M-H-90] − , [M-H-120] − , [M-H-180] − , and [M-H-210] − for pentosyl residues and [M-H-74] − and [M-H-104] − for deoxyhexosyl residues [41].

Table 1
Phytochemical screen and antioxidant activity of fruit, leaf, and root of C. pentagyna (TP: total phenol; TF: total flavonoid; TPA: total phenolic acid; TAC: total anthocyanin) [19]s with different superscript lowercase letters within the same row differ significantly (p < 0.05) quercetin, and isoquercetin in the Iranian C. pentagyna fruit extract.Moreover, a number of 39 compounds (flavonoid aglycones, flavonoid O-and C-glycosides, organic and phenolic acids and proanthocyanidins) were identified in Romania C. pentagyna leaf, flower and fruit ethyl acetate extracts[19].

Table 2
Phenolic constituents identified in the hydro-methanolic extracts of the fruit, leaf, and root of C. pentagyna using HPLC-ESI-MS/MS.Taleghani et al.BMC Complementary Medicine and Therapies (2024) 24:126

Table 3
Chemical composition of petroleum ether extracts from C. pentagyna fruit, leaf, and root analyzed by GC-MS.